CN101609093B - Test card for rapid diagnosis of pancreatictrauma and acute pancreatitis and test method thereof - Google Patents
Test card for rapid diagnosis of pancreatictrauma and acute pancreatitis and test method thereof Download PDFInfo
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- CN101609093B CN101609093B CN 200910060027 CN200910060027A CN101609093B CN 101609093 B CN101609093 B CN 101609093B CN 200910060027 CN200910060027 CN 200910060027 CN 200910060027 A CN200910060027 A CN 200910060027A CN 101609093 B CN101609093 B CN 101609093B
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Abstract
The invention relates to a test card for rapid diagnosis of pancreatictrauma and acute pancreatitis and test method thereof, aiming at solving the problems of low accuracy, long time for test, high device cost and inconvenient operation of the existing diagnosis method. A test line and a quality control line are arranged on a chromatography film, the test line is coated TAP polyclonal antibody, the quality control line is coated rabbit anti-mouse immunoglobulin antibody, and the concentration of the TAP polyclonal antibody and the rabbit anti-mouse immunoglobulin antibody is 1-100nmol/L.
Description
Technical field:
The present invention is with relevant with the apparatus and method of immunology diagnosis pancreatic trauma and acute pancreatitis.
Background technology:
Pancreatic trauma and acute pancreatitis are common clinically Acute critical diseases, and be if untimely treatment often jeopardizes patient's life, therefore extremely important for the life of saving the patient to the timely diagnosis of pancreatic trauma and acute pancreatitis.The routine diagnosis of pancreatic trauma and acute pancreatitis is mainly the amylase activity of measuring in peritoneal exudate and blood at present; with the sharply rising of its activity as diagnosis basis; but the accuracy of the method is not good enough, often causes and fails to pinpoint a disease in diagnosis with mistaken diagnosis and crisis patient life.In addition also by APACHE II scoring, pancreatic trauma and acute pancreatitis are diagnosed clinically, but this methods of marking is too loaded down with trivial details, if the scoring deficiency easily causes conditions of patients to be incured loss through delay; Too highly if mark easily cause overmedication, so this methods of marking can not satisfy the requirement to the quick diagnosis of pancreatic trauma and acute pancreatitis.In recent years, along with going deep into of research, discovery is when pancreatic trauma and acute pancreatitis generation, when activating, its critical event--trypsinogen produces a small-molecular peptides, be called trypsinogen activating peptide (TAP), the rising of this molecule content in ascites, blood and urine and the generation of pancreatic trauma and acute pancreatitis are closely related.By the TAP assay state of an illness of Diagnosis of Pancreatic wound and acute pancreatitis comparatively exactly not only, therefore can also carry out Efficient Evaluation to patient's prognosis, have the researcher begin this molecule as the specific diagnosis thing of pancreatic trauma and acute pancreatitis and obtain good effect.Mainly enzyme linked immunosorbent assay (ELISA) to the detection method of TAP at present, but the method grow (4～5 hours) consuming time, also need specific absorbance detection instrument and professional operation, price is also more expensive in addition, therefore can not satisfy the requirement to state of an illness quick diagnosis, more be not suitable for using at basic medical unit.
Summary of the invention:
The purpose of this invention is to provide a kind of easy to detectly, quick, highly sensitive, with low cost, be fit to the on-the-spot quick diagnosis pancreatic trauma of using and test card and the method for testing thereof of acute pancreatitis.
The object of the present invention is achieved like this:
The test card of quick diagnosis pancreatic trauma of the present invention and acute pancreatitis, p-wire and nature controlling line are arranged on chromatographic film, p-wire is coated TAP polyclonal antibody, nature controlling line is the coated anti-mouse immuning ball protein antibody of rabbit, and the concentration range of the anti-mouse immuning ball protein antibody of TAP polyclonal antibody and rabbit is 1-100nmol/L.
P-wire is three, and the spacing of line is 3-6mm, and the concentration of the TAP polyclonal antibody of 1-3 bar p-wire is followed successively by 1-10nmol/L, 10-50nmol/L, and 50-100nmol/L, nature controlling line are 1, and the spacing between the 3rd p-wire is 3-8mm.
Test card, the preparation method of TAP polyclonal antibody p-wire is as follows:
(1) synthetic TAP,
(2) preparation collaurum colloidal sol,
(3) preparation TAP monoclonal antibody,
A. adopt conventional glutaraldehyde cross-linking method, with TAP respectively with hemocyanin and crosslinked TPA-KLH conjugate and the TAP-BSA conjugate made of bovine serum albumin(BSA),
B. use TAP-KLH conjugate immunity Balb/c mouse,
C. get immune mouse spleen cell and commodity SP2/0 myeloma cell under polyglycol PEG-4000 effect, carry out Fusion of Cells according to conventional method, generate the hybridoma cell strain suspension,
D. adopt the hybridoma cell strain of the anti-TAP of indirect enzyme-linked immunosorbent assay screening,
E. hybridoma cell strain is inoculated in in the Balb/c mouse peritoneal of freund 's incomplete adjuvant after 1 week of pre-service, collection ascites adopts sad method or ammonium sulfate method or polyglycol PEG precipitation method purifying ascites namely to gather in the crops the TAP monoclonal antibody according to the difference of monoclonal antibody subclass.
(4) the TAP monoclonal antibody is added collaurum colloidal sol with the phosphate buffer dissolving as mother liquor, then goes namely to make again the TAP monoclonal antibody of colloid gold label with the phosphate buffer redissolution after supernatant after adding polyglycol PEG-10000 coreaction,
(5) with synthetic TAP as antigen, carry out hypodermic injection at the different parts of rabbit after preparing solution and mix with the Fu Shi Freund's complete adjuvant with phosphate buffer, collector's rabbit blood repeatedly, collect by hydro-extractor the antiserum that contains polyclonal antibody after removing blood clot, to namely get TAP Anti-TNF-α liquid solution after the antiserum purifying, with TAP Anti-TNF-α liquid solution make be coated on after variable concentrations on chromatographic film p-wire.
Test card is characterized in that the preparation method of the anti-mouse immuning ball protein antibody of rabbit nature controlling line is as follows:
The immunoglobulin (Ig) of Balb/c mouse as antigen, is carried out hypodermic injection to the rabbit different parts, repeatedly after collector's rabbit blood, centrifugal collection antiserum, with the antibody-solutions after purifying be coated on chromatographic film nature controlling line.
The method of testing of test card comprises the steps:
Patient's humoral specimen is directly splashed in sample well observe after placement 10min, if without the p-wire colour developing, be diagnosed as without applicable disease generation; If only article one p-wire colour developing is diagnosed as applicable disease may occur; If 1st, 2 p-wire colour developings are diagnosed as the applicable disease of very likely generation; If three p-wire all develops the color, be diagnosed as the applicable disease of severe; If nature controlling line without colour developing, shows test card and lost efficacy.
Test card of the present invention belongs to a kind of new specific diagnosis reagent, is mainly used in the quick diagnosis of pancreatic trauma and the acute pancreatitis state of an illness.This test card mainly utilizes the principle of antigen-antibody energy specific bond in immunology, utilize ripe colloidal gold immunochromatographimethod technology, with people source TAP as antigen, preparation detects the test card of TAP content, but the TAP in rapid semi-quantitative detection patient urine and peritoneal exudate reflects the PD degree of pancreatic trauma and acute pancreatitis, whole test process only needs 10～15 minutes, and have highly sensitive, high specificity, cheap and without any need for advantages such as supplementary instrument equipment, be highly suitable for that basic medical unit is on-the-spot to be used.
Description of drawings:
Fig. 1 is test card front view of the present invention.
Fig. 2 is the A-A cut-open view of Fig. 1.
Embodiment:
The structure of test card forms and processes
This test card is comprised of plastic casing 1 and test strips, its length is 80～110mm, and width is 20～35mm, wherein the plastic casing front have sample well 11 and as a result view window 10 be convenient to application of sample and observations, hand-held during the holding area convenient operation of afterbody, concrete structure is seen Fig. 1.Test strips is comprised of multilayer material, mainly comprises sample filtering layer 3, sample pad 4, pad 5, chromatographic film 6 (3 p-wires and 1 nature controlling line are arranged on film), adsorptive pads 7 and end liner 2.Selection material and the disposal route of each ingredient of test strips are as follows:
(1) sample filtering layer: formed by 3～5 metafiltration paper
(2) sample pad: made by all-glass paper, take out after the PBS immersion 2min with pH 7.0～8.4, below 80 ℃, oven dry or alternate manner are dry.
(3) pad: made by all-glass paper, take out after PBS immersion 2min with pH 7.0～8.4, oven dry below 80 ℃, be immersed in after cooling in the TAP monoclonal antibody solution of colloid gold label of 10～100nmol/L concentration range, take out at room temperature dry (4) chromatographic film after 10min: material is nitrocellulose filter (NC film), glutaraldehyde solution with 0.8% (g/ml) soaks 30min, take out, dry under room temperature, the TAP polyclonal antibody line of coated 3 variable concentrations is as detection line in the above, the coated 1 anti-mouse immuning ball protein antibody of rabbit line is as nature controlling line simultaneously.
Adsorptive pads: formed by 3～5 metafiltration paper or thieving paper
(5) end liner: scribble the non-absorbent material of adhesive sticker for one side, as the PVC plate
Adopt flush coater (commercially available) to be coated with the TAP polyclonal antibody on 3 detection lines, on 3 detection lines, the concentration of encrusting substance increases progressively successively, and its concentration range is respectively 1～10nmol/L, 10～50nmol/L, 50～100nmol/L; The coated scope of nature controlling line is the anti-mouse immuning ball protein antibody of rabbit of 1～100nmol/L.Article 3, the distance between detection line is 3～6mm, and the distance between the 3rd detection line and nature controlling line is 3～8mm.When 3 detection lines any one when presenting redness, the expression sample is positive; When the 1st colour developing of detection line, when the 2nd, 3 detection line do not develop the color, in the expression testing sample, the content of TAP was greater than 2nmol/L; When the colour developing of the 1st, 2 detection line, when the 3rd detection line do not develop the color, show that TAP content in testing sample is greater than 25nmol/L; When 3 detection lines all developed the color, in the expression testing sample, TAP content was greater than 90nmol/L; Nature controlling line develops the color all the time, does not show that test strips lost efficacy if do not develop the color.
Test philosophy
The principle of institute of the present invention foundation is: the TAP monoclonal antibody specificity of the colloid gold label of the TAP in sample to be checked on pad is combined, bond and being moved to the test paper other end under the promotion of chromatography effect by the monoclonal antibody of the moisture content dissolved gum body in sample gold mark, when moving to the detection line of coated TAP polyclonal antibody, form the double-antibody sandwich compound and assemble colour developing, and the TAP monoclonal antibody of free colloid gold label when moving to nature controlling line with the anti-mouse immuning ball protein antibody capture of rabbit and develop the color to show the validity of test card.
Application example
(1) sample liquid of preparation variable concentrations detects
1, with phosphate buffer (0.01mol/L, pH 7.0～7.5) compound concentration is that the TAP solution of 1nmol/L is as analyte sample fluid, get on 2～3 sample wells that directly drop in test card, observe after placing 10min, nature controlling line colour developing and detection line occur without obvious colour band, show that in sample, TAP content is lower than 2nmol/L.
2, with phosphate buffer (0.01mol/L, pH 7.0～7.5) compound concentration is that the TAP solution of 10nmol/L is as analyte sample fluid, get on 2～3 sample wells that directly drop in test card, observe after placing 10min, the 1st detection line and nature controlling line all develop the color.Show that in sample, TAP content is between 2～25nmol/L.
3, with phosphate buffer (0.01mol/L, pH 7.0～7.5) compound concentration is that the TAP solution of 50nmol/L is as analyte sample fluid, get on 2～3 sample wells that directly drop in test card, observe after placing 10min, 1st, 2 detection lines and nature controlling line all develop the color, and show that in sample, TAP content is between 25～90nmol/L.
4, with phosphate buffer (0.01mol/L, pH 7.0～7.5) compound concentration is that the TAP solution of 100nmol/L is as analyte sample fluid, get on 2～3 sample wells that directly drop in test card, observe after placing 10min, article 3, detection line and nature controlling line all develop the color, and show that in sample, TAP is greater than 90nmol/L.
(2) interference measurement
With normal person's urine and with the phosphate buffer of pH 7.0～8.4 be mixed with that concentration is 10,50, people's trypsase of 100nmol/L, human protease are former-1, human protease is former-2, human serum albumins, human immunoglobulin(HIg) solution, detect with test card, result is all negative, shows that above-mentioned substance does not all cause interference to test.
(3) patient specimen detects
1, urine specimen: get 2～3 patient urines and directly drop on the sample well of test card, observe after placing 10min, if without the detection line colour developing, show TAP content in urine lower than 2nmol/L, be diagnosed as without pancreas wound and acute pancreatitis and occur; If only the 1st detection line colour developing shows TAP content in urine between 2～25nmol/L, be diagnosed as pancreatic trauma and acute pancreatitis may occur; If only the 1st, 2 detection line colour developing shows TAP content in urine between 25～90nmol/L, be diagnosed as pancreatic trauma and acute pancreatitis very likely occur; If 3 detection line all develops the color, show that TAP content in urine higher than 90nmol/L, is diagnosed as severe pancreatic trauma and acute pancreatitis; If nature controlling line without colour developing, shows test card and lost efficacy.
2, peritoneal exudate sample: get 2～3 patient's peritoneal exudates and directly drop on the sample well of test card, observe after placing 10min, if without detection line colour developing, show TAP content in peritoneal exudate lower than 2nmol/L, be diagnosed as without pancreas wound and acute pancreatitis and occur; If only the 1st detection line colour developing shows TAP content in peritoneal exudate between 2～25nmol/L, be diagnosed as pancreatic trauma and acute pancreatitis may occur; If only the 1st, 2 detection line colour developing shows TAP content in peritoneal exudate between 25～90nmol/L, be diagnosed as pancreatic trauma and acute pancreatitis very likely occur; If 3 detection line all develops the color, show that TAP content in peritoneal exudate higher than 90nmol/L, is diagnosed as severe pancreatic trauma and acute pancreatitis; If nature controlling line without colour developing, shows test card and lost efficacy.
The preparation method for antibody of p-wire and the anti-line of matter is as follows:
1, people source TAP's is synthetic
Adopt the synthetic TAP molecule Ala-Pro-Phe-Asp-Asp-Asp-Asp-Lys (APFDDDDK) of polypeptide automatic synthesizer (production of U.S. BECKMAN company).
2, the preparation of collaurum colloidal sol is with gold chloride (HAuCL
4) be mixed with deionized water the solution that concentration is 1% (g/ml), be diluted to again 0.01% working fluid with deionized water, be heated to boiling, 1% (g/ml) trisodium citrate aqueous solution that adds again 15ml, continue to be heated to occur transparent orange red till, this solution is collaurum colloidal sol.
3, the preparation of TAP monoclonal antibody
(1) preparation of TAP conjugate: adopt conventional glutaraldehyde cross-linking method, TAP and hemocyanin (KLH) and bovine serum albumin(BSA) (BSA) is crosslinked, concrete grammar is: the TAP that 5mg is synthetic joins respectively in the KLH and BSA of 7mg, slowly add 1ml freshly prepared 0.3% (g/ml) glutaraldehyde solution while vibrating, react 2h under room temperature, with pH8.5 borate buffer dialysed overnight and get final product.The TAP-KLH conjugate of making and TAP-BSA conjugate are respectively used to mouse immune and monoclonal antibody screening;
(2) animal immune: with the TAP-KLH conjugate immunity Balb/C mouse in 6 ages in week, at first the TAP-KLH conjugate with 300 μ g carries out lumbar injection with the water-in-oil emulsion that Fu Shi Freund's complete adjuvant mixed in equal amounts becomes.Every 4 weeks, carry out lumbar injection with TAP-KLH conjugate and the freund 's incomplete adjuvant mixed in equal amounts of same dosage afterwards.Immunity is 4 times so altogether, and last 1 abdominal cavity and tail vein are respectively injected TAP-KLH conjugate 150 μ g and carried out booster immunization, and after 3 days, extracting spleen cell carries out Fusion of Cells;
(3) foundation of hybridoma cell line: get the SP2/0 myeloma cell of immune mouse spleen cell and purchase under polyglycol PEG-4000 (commercially available) effect, carry out Fusion of Cells according to conventional method.Cultivate with commercially available HAT nutrient culture media (cell culture medium that namely contains xanthine, aminopterin and thymidine), use commercially available employing HT nutrient culture media (cell culture medium that namely contains hypoxanthine and thymidine) after 1 week instead and cultivate;
(4) screening of TAP monoclonal antibody: adopt the monoclonal antibody hybridoma cell strain of the anti-TAP of indirect enzyme-linked immunosorbent assay screening secretion, concrete operations are: the TAP-BSA conjugate of 10 μ g/ml is coated in the hole of commercial polyethylene soft board according to a conventional method, adopt pH 9.6, the 0.02mol/L carbonate buffer solution is coated damping fluid.Add sample 100 μ l to be checked in coated good hole, hatch 1h for 37 ℃, washing, add goat-anti mouse enzyme labelled antibody (commercially available, working concentration is dilution in 1: 1000) 100 μ l, hatch 45min for 37 ℃, washing adds substrate 3 ', 3 ', 5,5 '-tetramethyl benzidine (TMB), 100 μ l are hatched 15min for 37 ℃, the sulfuric acid 100 μ l cessation reactions that add 2mol/L, colorimetric with the hybridoma cell strain of screening secretion TAP monoclonal antibody, is carried out cloning with limiting dilution assay under wavelength 450nm, carries out altogether 4 time clonings;
(5) ascites preparation and purification: with the hybridoma suspension inoculation in advance with the Balb/C mouse peritoneal after 1 week of freund 's incomplete adjuvant pre-service, collect ascites, namely gather in the crops the TAP monoclonal antibody according to the difference sad/ammonium sulfate method of employing or the polyglycol PEG precipitation method purifying ascites of monoclonal antibody subclass.
4, the colloid gold label of TAP monoclonal antibody
The TAP monoclonal antibody is dissolved and is diluted to 2～4mg/ml with phosphate buffer (0.01mol/L, pH 7.0～7.5), as mother liquor.Every 100ml collaurum colloidal sol adds 1～3ml antibody mother liquor, and vibration 2min is with the K of 0.2mol/L
2CO
3Regulate pH to 8.4, vibration 5min adds 2ml 11% (g/ml) polyglycol PEG-10000 (commercially available), vibration 5min, and the centrifugal 15min of 8000～15000rpm redissolves with phosphate buffer after removing supernatant.At the centrifugal 15min of 6000～13000rpm, remove supernatant, after being diluted with phosphate buffer, precipitation namely gets the TAP monoclonal antibody of colloid gold label.
5, the preparation of TAP polyclonal antibody
Synthetic people source TAP as antigen, is prepared concentration as the solution of 100 μ g/ml take aforementioned phosphate buffer, get the water-in-oil emulsion that 1ml and isopyknic Fu Shi Freund's complete adjuvant be mixed into and carry out hypodermic injection at 4 different parts of rabbit.Every 4 weeks repeat once, collector's rabbit blood after 4 times.Collect good blood and spend the night in 4 ℃ of placements after placing 30min at 37 ℃ again, remaining liq is transferred in plastic centrifuge tube after removing blood clot, under 4 ℃ 10, the centrifugal 10min of 000g collects supernatant and is the antiserum that contains polyclonal antibody.the purifying of antibody: adding solid sodium azide (commercially available) to form concentration in antiserum is the solution of 0.05% (g/ml), under 4 ℃ 15, the centrifugal 5min of 000g, the antiserum that shifts out clarification is used TBS damping fluid (6.06g Tris after being filtered and removing the unnecessary impurity such as fat, 8.78g NaCl and 0.5g sodium azide are dissolved in 1L distilled water, and regulate pH 7.4 with HCl) with 1: 5 dilution proportion, refilter, then with the speed of per minute 0.5ml, antiserum is injected albumin A sepharose CL-4B affinity chromatographic column (commercially available), with TBS buffer solution elution affinity post, and collect eluent and detect it in the absorbance log of 280nm, (the 3.75g glycocoll is dissolved in 1L distilled water to add elution buffer after the absorbance log of eluent is less than 0.008 again, regulate pH 2.7 with HCl), being eluted to all albumen with the speed of 0.5ml/min all flows down.With adding 100ul neutralization buffer solution (121.2g Tris, 87.8g NaCl, 0.37g EDTA and 5g sodium azide are dissolved in 1L distilled water, and regulate pH8.0 with HCl) 1.5ml EP pipe be in charge of the collection eluent, pH checks the pH of eluent after mixing with the pH test paper, if can utilize neutralization buffer to transfer to approximately pH7.4 to prevent the sex change of antibody lower than 7.Add 10ml in post, pHl.9 elution buffer solution, collect as stated above eluent to absorbance less than 0.008, the Anti-TNF-α liquid solution that is purifying of collecting in all EP pipes.
6, the preparation of the anti-Balb/C mouse immuning ball protein of rabbit antibody
The immunoglobulin (Ig) of Balb/C mouse as antigen, is carried out immunity to rabbit, and collection and the purifying of antibody are the same.
Claims (2)
1. the test card of quick diagnosis pancreatic trauma and acute pancreatitis, it is characterized in that p-wire and nature controlling line are arranged on chromatographic film, p-wire is coated trypsinogen activating peptide polyclonal antibody, nature controlling line is the coated anti-mouse immuning ball protein antibody of rabbit, the concentration range of the anti-mouse immuning ball protein antibody of trypsinogen activating peptide polyclonal antibody and rabbit is 1-100nmol/L
Described p-wire is three, the spacing of line is 3-6mm, the concentration of the trypsinogen activating peptide polyclonal antibody of 1-3 bar p-wire is followed successively by 1-10nmol/L, 10-50nmol/L, 50-100nmol/L, described nature controlling line is 1, and the 3rd the spacing between p-wire is 3-8mm, and the preparation method of trypsinogen activating peptide polyclonal antibody p-wire is as follows:
(1) trypsin biosynthesis pepsinogen activating peptide,
(2) preparation collaurum colloidal sol,
(3) preparation trypsinogen activating peptide monoclonal antibody,
A. adopt conventional glutaraldehyde cross-linking method, with the trypsinogen activating peptide respectively with hemocyanin KLH and crosslinked trypsinogen activating peptide-KLH conjugate and the trypsinogen activating peptide-BSA conjugate made of bovine serum albumin BSA,
B. use trypsinogen activating peptide-KLH conjugate immunity Balb/c mouse,
C. get immune mouse spleen cell and commodity SP2/0 myeloma cell under polyglycol PEG-4000 effect, carry out Fusion of Cells according to conventional method, generate the hybridoma cell strain suspension,
D. adopt the hybridoma cell strain of indirect enzyme-linked immunosorbent assay screening trypsinogen activation peptides,
E. hybridoma cell strain is inoculated in in the Balb/c mouse peritoneal of freund 's incomplete adjuvant after 1 week of pre-service, collect ascites and namely gather in the crops trypsinogen activating peptide monoclonal antibody according to difference employing sad method/ammonium sulfate method or the polyglycol PEG precipitation method purifying ascites of monoclonal antibody subclass
(4) trypsinogen activating peptide monoclonal antibody is added collaurum colloidal sol with the phosphate buffer dissolving as mother liquor, add again and go after polyglycol PEG-10000 coreaction after supernatant to redissolve with phosphate buffer the trypsinogen activating peptide monoclonal antibody of namely making colloid gold label, standby
(5) with synthetic trypsinogen activating peptide as antigen, carry out hypodermic injection at the different parts of rabbit after preparing solution and mix with the Fu Shi Freund's complete adjuvant with phosphate buffer, collector's rabbit blood repeatedly, collect by hydro-extractor the antiserum that contains polyclonal antibody after removing blood clot, trypsinogen activating peptide Anti-TNF-α liquid solution will namely be got after the antiserum purifying, with trypsinogen activating peptide Anti-TNF-α liquid solution make be coated on after variable concentrations on chromatographic film p-wire, the structure of described test card is as follows: chromatographic film (6) is positioned at shell (1), under end liner (2) is arranged, p-wire on chromatographic film (6) and the nature controlling line transparent windows (10) on the shell, the p-wire left side has pad (5) to be connected with sample pad (4), filtering layer (3) is arranged on sample pad (4), sample well (11) is arranged on the shell on filtering layer (3), there is adsorptive pads (7) on the right side of nature controlling line, pad (5): made by all-glass paper, take out after PBS immersion 2min with pH7.0-8.4, oven dry below 80 ℃, be immersed in after cooling in the trypsinogen activating peptide monoclonal antibody solution of colloid gold label of 10-100nmol/L concentration range, take out at room temperature dry after 10min.
2. test card according to claim 1 is characterized in that the preparation method of the anti-mouse immuning ball protein antibody of rabbit nature controlling line is as follows:
The immunoglobulin (Ig) of Balb/c mouse as antigen, is carried out hypodermic injection to the rabbit different parts, repeatedly after collector's rabbit blood, centrifugal collection antiserum, with the antibody-solutions after purifying be coated on chromatographic film nature controlling line.
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| CN102879583B (en) * | 2012-09-21 | 2014-11-19 | 任建东 | Test card for rapidly diagnosing pancreatic trauma and acute pancreatitis |
| WO2014199954A1 (en) * | 2013-06-10 | 2014-12-18 | 旭化成せんい株式会社 | Immunochromatographic diagnosis kit |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4948723A (en) * | 1987-01-16 | 1990-08-14 | Bioscience International, Inc. | Method of diagnosis and severity-assessment of pancreatic disease |
| CN1746675A (en) * | 2004-09-07 | 2006-03-15 | 李人 | Immune chromatograph testing strip and production thereof |
| CN101067627A (en) * | 2007-06-14 | 2007-11-07 | 中国农业科学院兰州兽医研究所 | Asial type foot-and-mouth disease virus fast diagnose test paper bar and producing method thereof |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4948723A (en) * | 1987-01-16 | 1990-08-14 | Bioscience International, Inc. | Method of diagnosis and severity-assessment of pancreatic disease |
| CN1746675A (en) * | 2004-09-07 | 2006-03-15 | 李人 | Immune chromatograph testing strip and production thereof |
| CN101067627A (en) * | 2007-06-14 | 2007-11-07 | 中国农业科学院兰州兽医研究所 | Asial type foot-and-mouth disease virus fast diagnose test paper bar and producing method thereof |
Non-Patent Citations (3)
| Title |
|---|
| Hurley PR, Cook A, Jehanli A, Austen BM, Hermon-Taylor J.Development of radioimmunoassays for free tetra-L-aspartyl-L-lysine trypsinogen activation peptides (TAP).《Journal of Immunological Methods》.1988,第111卷(第2期),195–203. * |
| JP Neoptolemos et al..Early prediction of severity in acute pancreatitis by urinary trypsinogen activation peptide: a multicentre study.《The Lancet》.2000,第355卷(第9219期),1955–1960. * |
| Tenner S et al..Urinary trypsinogen activation peptide (TAP) predicts severity in patients with acute pancreatitis.《Int J Pancreatol》.1997,第21卷(第2期),105-110. * |
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